The long term objective of the research program is the understanding of nervous system function as influenced by benzodiazepines and opiates. The immediate aims of the benzodiazepine research are: 1) production of monoclonal antibody to benzodiazepine/GABA/chloride ion channel complex proteins which have been purified using a ligand affinity method. 2) application of these antibodies to purify the individual components of this receptor complex. 3) complete purification of the receptor proteins using HPLC. 4) reconstitution of the receptor complex proteins into nerve cells. 5) use of antisera prepared against the receptor complex to study the neurophysiology and metabolism of this structure in neurons. The significance of this work is derived from the role of this receptor/ion channel as a prominent and widely distributed inhibitory conductance mechanism in the central nervous system. Further, it is the site of action of clinically important drugs, such as the benzodiazepines (i.e., Valium) and barbiturates. With respect to the study of opiate neurobiology, this research will focus upon the potential interaction between opioid peptides and the (putative) acidergic neurotransmitters, glutamate and aspartate, in spinal cord and sensory neurons. This will be achieved with the application of antisera prepared against the amino acids and the enzymes, glutaminase and aspartate aminotransferase, which may serve as markers of acidergic neuronal function. The synthesis, cellular release and immunohistochemical distribution of glycylglutamine, a dipeptide derived from beta-endorphin, will also be studied.